tbuckley/cabot-search huggingface dataset:
Abstract P1086: Lrrc10 Is A Multifaceted Modulator Of Cardiac Calcium And Sodium Channels
Authors: Pedro J Del Rivero Morfin, Nourdine Chakouri, Manu Ben Johny
Journal: Circulation Research (2022)

L-type calcium channels (LTCC) are vital for heart physiology as they allow entry of calcium to the cardiomyocyte, ultimately leading to contraction. These channels are exquisitely modulated by auxiliary subunits and a vast array of regulatory proteins. Heart-specific leucine-rich repeat-containing protein 10 (Lrrc10) plays a key role as a regulator of LTCC. Previous work has shown that Lrrc10 knockdown in zebrafish disrupts development of the cardiac tube, while ablation of Lrrc10 in mice results in heart dysfunction and maladaptation to increased cardiac afterload. Moreover, human mutations in Lrrc10 gene have been linked to dilated cardiomyopathy, and sudden unexplained nocturnal death syndrome (SUNDS). Still, the molecular mechanisms underlying Lrrc10 regulation of LTCC is not fully established. Beyond this, it is unknown whether Lrrc10 modulates other cardiac ion channels. Here, we hypothesized that Lrrc10 serves as a versatile regulator of four-domain voltage-gated ion channels, while mutations in Lrrc10 differentially disrupt channel regulation. Using whole-cell and single-channel electrophysiology, we found that Lrrc10: 1) upregulates LTCC currents by increasing peak open probability; 2) accelerates calcium-dependent inactivation; and 3) evokes a hyperpolarizing-shift in voltage-dependence of activation. Flow cytometric FRET 2-hybrid assay showed that Lrrc10 interacts with LTCC cytoplasmic subunits, specifically the N- and C-termini. We further used FRET 2-hybrid analysis to evaluate the interaction of Lrrc10 with related cardiac ion channels. We observed robust Lrrc10 interaction with Na V 1.5 channels. Multi-channel electrophysiology recordings revealed increased late sodium currents, suggesting that Lrrc10 also modulates Na V 1.5 activity. Additionally, electrophysiological studies showed that Lrrc10 variants linked to dilated cardiomyopathy and SUNDS disparately alter modulation of calcium and sodium channels. In conclusion, Lrrc10 is a multifaceted regulator of four-domain, voltage-gated cardiac ion channels, a finding that will lend critical insights into heart function and disease.
Late onset asthma?
Authors: RM Aleva, B Wouters, J Kraan, PE Postmus
Journal: European Respiratory Journal (1991)

A 77 yr old male was referred for dyspnoea and recurrent infections of the lower airways for the previous five months. On the lateral chest roentgenogram a process in the tracheal area was found, and during bronchoscopy a polypoid tumour was seen about four centimetres above the main carina. It appeared to be a benign endotracheal tumour which was removed after coagulation by a Neodymium yttrium aluminium garnet (YAG) laser.
Abstract P2100: Scalable Ultra-purification Of Adeno-associated Viral Vectors - A Novel Standard To Boost Transduction Efficacy And Potency For Cardiac Gene Therapy Development
Authors: Andreas Jungmann, Stephanie Simon, Philipp Schlegel, Eric Meinhardt, Charlotte Steeg, Jasmin Kroemer, Thomas Ruppert, Karsten Richter, Michelle Nessling, Philip W Raake, Norbert Frey, Patrick Most, Hugo A Katus, Martin Busch
Journal: Circulation Research (2022)

Introduction: A high transduction efficacy and potency of AAV-based cardiac gene therapies is key for the clinical translation in which hereditary and acquired cardiac disorders will be targeted. As such, ultrapure vectors with superior biological and therapeutic capabilities are a must for these therapies. Hypothesis: We hypothesized that our developed affinity chromatography (AC) based purification system will increase AAV recovery and - potency. Methods: AAV vector production, AC based - or iodixanol density gradient (DG) based purification, Q-PCR, WB, EM, LC-MS/MS, P-loop Results: Using the same vector input quantity, the AC-based purification enabled an approximately 13-fold greater vector genome copy (vgc) recovery than the DG-based purification. Mass spectrometry analysis demonstrated ultrapure AAV9 vectors as a result of an AC purification (AAV9 93.24% vs. 6,76% contaminants) whereas corresponding DG preparations resulted in highly contaminated vectors (AAV9 5.49% vs. 94,51% contaminants). Biological potency of AC- and DG-purified AAV9 vectors towards cardiac transduction were determined by systemic injections of 1·10 10 , 1·10 11 or 1·10 12 vgc of AAV9-EGFP in C57BL/6 mice (n=8 each group). AC-purified AAVs achieved a significantly higher cardiac transduction efficacy for every dosage assessed by comparative bulk myocardial DNA, RNA and protein level analysis after 2 weeks. Therapeutic potency was examined for a recently published novel target for chronic heart failure namely the RFXP1-RLN system. To this end, a dosage of 5·10 11 vgcs of either AC- or DG purified AAV9-RXFP1 vectors were systemically injected and the cardiac contractile performance increase was captured after 2 weeks in mice of both groups. Of note, 10 minutes after RLN administration, the rise in LV +dp/dt max. was already significantly greater in the AC- than the DG-vector treated group (AC: 13594+/-1972 vs. DC: 9822+/-801 mmHg/s; n=8 per group, p&lt;0.01). Conclusion: The data clearly promote AC-based AAV purification as a novel standard for cardiovascular basic and translational research. Higher consistency in results, higher therapeutic effects and superior biological potency can be expected from higher production yields of ultrapure AAVs even at lower vector dosages.
Abstract P3121: Serological Mechanisms Driving Cardiotoxicity In Covid-19
Authors: Maria T Colzani, Johannes Bargehr, Federica Mescia, Vincent Knight-Schrijver, Charlotte Summers, Kenneth G Smith, Paul Lyons, Sanjay Sinha
Journal: Circulation Research (2022)

Cardiac involvement is common in patients severely ill with COVID-19 and is associated with increased morbidity and mortality. The mechanism for myocardial injury in COVID-19 has been proposed to be at least in part due to a direct viral cytotoxic effect on cardiomyocytes. Preclinical data corroborate the notion that SARSCoV-2 can enter and replicate within hiPSC-derived cardiomyocytes. However, clinical evidence demonstrating direct viral effects on the heart remains limited. Alternatively, cardiac injury may result from excessive inflammatory cytokine production which is the main driver for the pathogenesis in COVID-19. Despite this, cardiotoxicity secondary to hyperinflammation constitutes a potentially amenable mechanism that remains largely unexplored. To investigate serological drivers of cardiotoxicity in COVID-19 we have established a bioassay that assessed the effects of serum from COVID-19 confirmed patients on human embryonic stem cell-derived cardiomyocytes (hESC-CMs). We demonstrate that serum from COVID-19 positive patients significantly reduced cardiomyocyte viability and contractility independent of viral transduction. Serum from patients with greater disease severity led to worse cardiomyocyte viability and this significantly correlated with levels of key inflammatory cytokines, including IL-6, TNF-α, IL1-β, IL-10, CRP and neutrophil to lymphocyte ratio with a specific reduction of CD4+ and CD8+ cells. Combinatorial blockade of IL-6 and TNF-α partially reversed the phenotype and preserved cardiomyocyte viability and function. Interestingly, treatment with serum from Systemic Inflammatory Response Syndrome (SIRS) patients recapitulates the effect observed with COVID-19 patients’ serum indicating that the phenotype observed is most likely not unique to COVID-19 disease. In conclusion, our results provide direct evidence that inflammatory cytokines are at least in part responsible for the cardiovascular damage seen in COVID-19 and indicate possible targets for therapeutic intervention.
Abstract P3059: Hydrogen Sulfide Inhibits Aortic Valve Fibrosis, Inflammation And Calcification By Inducing Pro-autophagy Effect In Human Valvular Interstitial Cells
Authors: Naaleum Song, Eunhye Ji, Kyoung-Hee Choi, Jeong-Eun Yu, Sahmin Lee
Journal: Circulation Research (2022)

Background: As of now, there is no pharmacological treatment for aortic valve stenosis (AS) except for valve replacement. In the previously studied proteomics analysis, we demonstrated that the expression of sulfide:quinone oxidoreductase (SQOR) was significantly lower in aortic valve interstitial cells (VICs) isolated from AS patients than those of normal patients. However, the role of SQOR in AS pathophysiology has not been clarified yet. Methods: To elucidate the cellular mechanism of SQOR and hydrogen sulfide (H2S) as therapeutic aspects in AS, we studied the effects of NaHS (H2S-releasing compound) on diseased aortic VICs using real-time PCR and western blots. Results: NaHS treatment led to increased SQOR expression in diseased aortic VICs and induced an increase of the Nrf2-target genes, especially NQO1. It showed that the effects of NaHS decreased not only fibrosis and inflammation markers (MMP9, TNF-α, IL6) but also calcification markers (ALP, osteocalcin, RUNX2, COL1A1). In addition, AMPK and Beclin-1 significantly increased by NaHS while Akt and mTOR decreased which is expected to have pro-autophagy effect. Conclusion: H2S releasing compound enhances the expression of SQOR and activates AMPK/mTOR mediated pro-autophagy function resulting in inhibition of fibrosis, inflammation and calcification in human VICs. Therefore, H2S can be a potential therapeutic target in AS.
Abstract P2059: Poly(rC)-binding Protein-1 (Pcbp1) Is Essential For Heart Development
Authors: Yao Wei Lu, Zhuomin Liang, Haipeng Guo, Tiago Fernandes, Xiaoyun Hu, Ramon Espinoza-Lewis, Douglas B Cowan, Hong Chen, Da-Zhi Wang
Journal: Circulation Research (2022)

Rationale: Proper development of the heart relies on a tightly regulated, yet diverse, pattern of gene expression that is governed by transcriptional, post-transcriptional, and translational processes. Congenital heart disease (CHD) is the most common birth defect and a leading cause of morbidity and mortality in children. While the genetic cause of some types of CHD has been identified, the molecular basis for the rest remains elusive. Poly(rC)-binding protein 1 (Pcbp1) is a RNA-binding protein that regulates RNA processing as well as post-transcriptional and translational processes in a variety of biological systems. Hypothesis: Pcbp1 play a critical role in regulating heart development by governing Notch and UPR pathways and mediating proper Aars2 gene splicing. Methods and Results: Germline deletion of Pcbp1 results in lethality before embryonic day (E) 8.5. We generated a cardiac-specific deletion of Pcbp1 by crossing Pcbp1-Flox with cTNT-Cre mice (Pcbp1-cKO cTNT ) and found that Pcbp1-cKO cTNT mice is 50% lethal perinatally. Embryonic hearts of Pcbp1-cKO cTNT mice displayed ventricular non-compaction and abnormal ventricular apex formation. Deep RNA sequencing of Pcbp1-cKO cTNT hearts revealed alteration of gene expression profiles reflective on ventricular maturation delay and dysregulation of Notch and UPR pathways. Interestingly, loss of Pcbp1 in cardiomyocytes disrupts alternative splicing of many important genes including Aars2, a gene associated with congenital mitochondrial cardiomyopathy. Pcbp1 deficiency resulted in creation of an Aars2 exon16-skipping variant, leading to its premature termination. eCLIP-seq showed that Pcbp1 primarily binds to CU-rich motifs at 3’UTR, distal intron and CDS regions of targets, and it interacts with regions of Aars2 transcript near exon 16. Using CRISPR/Cas9 technology, we knocked in loxP sites flanking the exon 16 of Aars2 (Aars2-Flox), and cross the floxed mice with cTNT-Cre mice to generate cardiac-specific exon16 deletion mutant of Aars2 (Aars2-cKO cTNT ). Intriguingly, abnormality in Aars2-cKO cTNT embryonic heart phenocopy aspects of ventricular non-compaction and malformation observed in Pcbp1-cKO cTNT heart. Accordingly, the transcriptome from hearts of Pcbp1-cKO cTNT and Aars2-cKO cTNT display high concordance and similarity and share strikingly common dysregulated pathways. Conclusions: We discover a novel function of Pcbp1 in heart development by regulating Notch and UPR pathways. Additionally, Pcbp1 is indispensable for Aars2 gene splicing, whose deficiency is associated with congenital cardiomyopathy. Our findings suggest modulating Pcbp1 in developing hearts may offer a novel therapeutic intervention for congenital heart failure.
Allergic alveolitis following exposure to epoxy polyester powder paint containing low amounts (&lt;1%) of acid anhydrides
Authors: P Piirila, H Keskinen, S Anttila, M Hyvonen, P Pfaffli, T Tuomi, O Tupasela, M Tuppurainen, H Nordman
Journal: European Respiratory Journal (1997)

Only one case report concerning allergic alveolitis caused by polyester powder paint has been published previously. The aim of this study was to determine whether phthalic anhydride (PA) or trimellitic anhydride (TMA) is the alveolitis-causing agent in such paint. A 61 year old woman showed recurrent symptoms of chills, cough, and fever whilst at work. She was working in a plant where epoxy polyester powder paints were used to paint metal. The paint was found to contain low (&lt;1%) amounts of TMA and PA. The patient showed shadowing on chest radiographs. In bronchoalveolar lavage, lymphocytosis (67%) and a low T-helper/T-suppressor ratio (0.2) were found. Transfer factor was within normal limits, but a slight reduction was verified after re-exposure to the paint. The symptoms, exposure, reduction in transfer factor, findings on chest radiographs and bronchoalveolar lavage were consistent with allergic alveolitis. In conclusion, the polyester powder paint used in the plant caused allergic alve olitis in this patient. Of the constituents in the paint, trimellitic anhydride and phthalic anhydride were the possible causative agents.
Abstract P2065: Ischemic Injury Aggravated Muscle-specific Mir-499-5p-impaired Angiogenic Property Of Endothelial Cell In Hindlimb Of Diabetic Db/db Mice: Role Of Small Extracellular Vesicles
Authors: Zhongjian A Cheng, Vandana Mallaredy, MAY TRUONGCAO, Chunlin Wang, Carolina Gonzalez, Maria Cimini, Grace Huang, Anna Maria Lucckese, Jessica Ibetti, Cindy Benedict, Sudarsan Rajan, Venkata Garikipati, Suresh Verma, Walter J Koch, Raj Kishore
Journal: Circulation Research (2022)

Background: Recently, skeletal muscle cells (SKMCs) have been reported to be critical for regulation of EC function in limbs. miR-499, a muscle specific microRNA (miR), was found to be modulated in diabetes and ischemic injury. Here, we studied the role of miR-499 in EC dysfunction in diabetic ischemic limb injury. Methods: ECs and SKMCs were isolated from ischemic or non-ischemic hindlimb (IHL) of male db/+ and db/db mice. Serum- and SKMC-derived small extracellular vesicles/exosomes (EV/Exo) were isolated by gradient ultracentrifugation. Ischemic hindlimb (IHL) surgery was conducted by unilateral ligation of formal artery. Results: miR-499-5p level was increased in SKMCs and ECs of db/db mice which was synergistically increased by ischemic injury. Overexpression of miR-499-5p impaired tube formation and migratory activity of ECs. Intramuscular injection of anti-miR-499-5p lentivirus improved blood prefusion and neovascularization in IHL of db/db mice. Mechanistically, miR-499-5p level was enhanced in serum- and SKMC-derived EV/Exo from db/db mice which was synergistically increased by ischemic injury. Diabetic SKMC-EV/Exo impaired blood perfusion in wildtype mice. Anti-miR-499-5p rescued diabetic SKMC-EV/Exo-impaired EC function. Co-culture of diabetic SKMCs with wildtype ECs increased miR-499-5p expression in ECs which was inhibited by EV/Exo inhibitor GW4869. Sex-determining region Y-box 6 (SOX6), the most attractive gene targeted by miR-499-5p, was decreased in ECs from db/db mice which was synergistically reduced by ischemic injury. SOX6 siRNA impaired pro-angiogenic factor secretion and function of ECs. Anti-miR-499-5p significantly enhanced SOX6 level in SKMs from IHL of db/db mice. Finally, overexpression of SOX6 by transduction of lentivirus improved EC function of db/db mice. Conclusions: Enhanced miR-499-5p expression in ECs of SKMs from hindlimb of db/db mice is synergistically increased by ischemic injury. EV/Exo transfer miR-499-5p from SKMCs to ECs. miR-499-5p impairs angiogenic property of EC via, at least partially, SOX6/proangiogenic factors axis. miR-499-5p may be a novel target for treatment of critical limb ischemia in diabetic patients.
Abstract P3124: Treatment Of Apoe <sup>-/-</sup> Mice With Advanced Atherosclerosis With The Senolytic Agent Abt-263 Caused Increased Mortality And Was Associated With Increased Endo-mt But Reduced Acta2 Cap Thickness And Collagen Content
Authors: Santosh Karnewar, Vaishnavi Karnewar, Gary K Owens
Journal: Circulation Research (2022)

Background: Plaque rupture and erosion of advanced atherosclerotic lesions resulting in ischemic heart disease or stroke are the leading worldwide cause of death. Multiple studies have shown that senescent cells are present in human atherosclerotic lesions, and recent animal studies have claimed that removing senescent cells from the lesions using senolytic agents has some beneficial effects but also limitations. Although these mouse studies have important implications for using senolytic agents to treat atherosclerosis, due to the lack of lineage tracing models and relying on markers not specific for various lesion cell types, it is difficult to ascertain the mechanisms that underline the beneficial effects shown by previous studies. Hypothesis: Treatment of western diet (WD) fed Apoe -/- mice with the senolytic agent ABT-263 (Navitoclax) will reduce lesion size and induce changes consistent with increased plaque stability including a thicker collagen-rich ACTA2 fibrous cap. Methods: SMC (Myh11-ERT2-Cre-eYFP+) and EC (Cdh5-ERT2-Cre-eYFP+) lineage tracing Apoe -/- were fed a WD for 18 weeks, followed by ABT-263 (100MG or 50MG /KG/BW) treatment for six to nine weeks on WD and analyzed for the indices of stability. Results: ABT-263 treatment did not result in changes in lesion size, lumen area, or outward remodeling of the brachiocephalic artery (BCA). However, surprisingly, ABT-263 treatment was associated with an 80% reduction in SMC (Myh11-eYFP+) but a 50-60% increase in endothelial cell (EC) contributions to lesions via EC to mesenchymal transition (EndoMT) (Chd5-eYFP+). ABT-263 treatment also reduced ACTA2 cap thickness by 50-75% and collagen content by 50%, but increased mortality by 50-60%. Conclusions: Taken together, contrary to our hypothesis, the senolytic agent ABT-263 treatment on advanced atherosclerosis showed multiple detrimental effects, including reduced indices of stability, increased Endo-MT, and increased mortality.
Abstract P1037: Polycythemia Vera Is Not Associated With Higher Inpatient Mortality In Patients Hospitalized For Myocardial Infarction: A Nationwide Analysis
Authors: Richard Orji, Emmanuel Ukenenye, Shirly Samuel, Mariam Khabsa, Gizem Yassa, Saad Waraich, Maryna shayuk
Journal: Circulation Research (2022)

Introduction: Polycythemia vera (PV) is a well-known myeloproliferative disorder with an abundance of red blood cells as well as white blood cells and platelets. This condition is also known as hyperviscosity syndrome (HVS). Myocardial infarction (MI), a lethal disease, occurs due to impaired coronary blood supply to the heart. The hyperviscosity of coronary arterial blood flow in PV patients is associated with an increased risk for coronary syndrome. However, our study sought to determine if PV worsens the outcomes or aggravates ongoing MI in hospitalized patients. Methods: We queried the National Inpatient Sample (NIS) database from 2016 to 2019. The NIS is the largest inpatient hospitalization database in the United States. The NIS was searched for hospitalization of adult patients with MI as a principal diagnosis with and without PV as a secondary diagnosis using ICD-10 codes. The primary outcome was inpatient mortality. The secondary results were Acute kidney injury (AKI), Cardiogenic shock (CS), Cardiac arrest (CA), Total hospital charge (THC), and length of stay (LOS). Multivariate logistic and linear regression analysis were used accordingly to adjust for confounders. Results: About 2,622,939 patients were admitted for MI; 0.091% (2,385) had underlying PV. Cohorts with PV vs No PV had a mean age of 69.2 years [CI 68.0 - 70.5] vs 66.9 years [CI 66.9 - 67.0]; male (66.2% vs 62.3%), female (33.8% vs 37.7%); white (85.3% vs 73.4%), black (5.6% vs 11.4%), and Hispanic (5.0% vs 8.8%). Compared to patients without PV, patients admitted with coexisting PV had similar inpatient mortality (5.7% vs 4.6%, AOR 1.17, 95% CI 0.79 - 1.73, P 0.437), AKI (20.3% vs 19.9%, P 0.546), CS (8.0% vs 6.3% P 0.222), CA (2.5% vs 2.9% P 0.690), THC (IRR 1.05, 95% CI 0.95 - 1.17, P 0.342), and LOS (IRR 1.06, 95% CI 0.95 - 1.18, P 0.277). Conclusion: Patients admitted primarily for MI with co-existing polycythemia vera had similar inpatient mortality. There was no difference in AKI, CS, CA, THC, and LOS in both groups. Phlebotomy in hospitalized Patients with MI and co-existing PV should be carried out with caution due to the risk of hypovolemia and possibly exacerbating the infarct. Additional studies are required to elaborate on the delicate management of this patient group.
Abstract P3108: Induced-Pluripotent Stem Cell-Derived Cardiomyocytes As A Model To Investigate Cardiotoxicity Of A Panel Of Breast Cancer Drugs
Authors: Elizabeth R Matthews, Omar Johnson, Jose A Gutierrez, Michelle Ward
Journal: Circulation Research (2022)

Anthracyclines (AC) are a class of antineoplastic drugs which are used to treat breast cancer (BC) subtypes including triple-negative BC. The 2019 Surveillance, Epidemiology, and End Results study indicates high BC survival rates and low reoccurrence with chemotherapy regimens that include AC. AC treatment, however, is associated with developing cardiovascular disease (CVD) in a subset of people, despite these patients having no history of CVD. Because CVD is typically irreversible, understanding patient susceptibility to AC toxicity is critical to cancer treatment management and long-term patient survival. As a first step towards understanding the genetic and mechanistic basis for AC-induced cardiotoxicity, we have developed an in vitro cardiomyocyte (CM) model using a panel of four genotyped induced-Pluripotent Stem Cell (iPSC) lines to test the effects of three AC (Doxorubicin, Epirubicin and Daunorubicin) and two non-AC (Mitoxantrone and Trastuzumab) BC drugs. We can reliably differentiate beating CMs at a median purity 90.5% based on the expression of cardiac-specific troponin T. Drug exposure for 48 hours reveals cardiotoxicity at micromolar concentrations across all drugs except for Trastuzumab, with Daunorubicin and Mitoxantrone exhibiting the greatest effect. We observe mRNA expression changes of stress response genes such as p53 within six hours of drug treatment, suggesting that an early transcriptional response mediates these effects. Using this model, we plan to measure the global transcriptional response to drug exposure across individuals and correlate these changes with each individual’s genetic variants to gain insight into cardiotoxicity susceptibility.